LOCUS       sequence-365751-        8940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the Z18 helixCAM 
            and constitutive expression of an identifying fluorescent protein 
            (eGFP). Flanked with PiggyBac sites for easy line construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8940)
  AUTHORS   Chao G, Wannier TM, Gutierrez C, Borders NC, Appleton E, Chadha A, 
            Lebar T, Church GM
  TITLE     helixCAM: A platform for programmable cellular assembly in bacteria 
            and human cells.
  JOURNAL   Cell. 2022 Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012.
  PUBMED    36055250
REFERENCE   2  (bases 1 to 8940)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2022 
            Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012."
COMMENT     Sequence Label: PB_TRE_Z18-PDGFR_EF1a_eGFP
FEATURES             Location/Qualifiers
     source          1..8940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(44..148)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(174..629)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(311..332)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     523..542
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     756..778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     770..787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     771..787
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   917..979
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1428..1806
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1436..1454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1472..1490
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1508..1526
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1544..1562
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1580..1598
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1616..1634
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1652..1670
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1858..1896
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1941..1965
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     1965..2024
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2028..2054
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2196..2225
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2229..2375
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(2378..2402)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2407..2448
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2462..2479
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2536..2553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2542..2749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2826..3037
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3050..3318
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3352..4095
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4096..4149
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4150..4152,4153..4155,4156..4866)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4150..4152,4153..4155,4156..4866)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4195..4216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4456..4475)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4803..4824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4867..4920
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4921..5520
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4921..4940)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5417..5437
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5553..5674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5590..5609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5644..5663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6437..6471
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6962..6978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6962..6978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6975..6997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6986..7002
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7010..7016,7017..7034,7035..7040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7055..7076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7193..7210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7364..7952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7444..7463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8123..8914,8915..8940,1..43))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8746..8765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
       61 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
      121 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
      181 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
      241 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
      301 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
      361 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
      421 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
      481 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
      541 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
      601 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
      661 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      721 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      781 ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg ggcagactcg
      841 cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca cgctgcagaa
      901 cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
      961 tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
     1021 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
     1081 tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct tctataaagt
     1141 aacaaaactt ttatgaggga cagccccccc ccaaagcccc cagggatgta attacgtccc
     1201 tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc
     1261 cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat
     1321 cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg
     1381 ggaaaaggcc tccacggcca ctagtctttc gtcttcaaga attcctcgag tttactccct
     1441 atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag
     1501 actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga
     1561 gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct
     1621 atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc
     1681 tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac cgtcagatcg
     1741 cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac ttcctaccct
     1801 cgtaaaggtc tagagctagc gaattcgaag ccaccaactg gcaggtgcgt atgtcgaggt
     1861 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     1921 tggagaattg gctagcatca acaagtttgt acaaaaaagt tggcatggag acagacacac
     1981 tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgactat ccatatgatg
     2041 ttccagatta tgctggggcc tcaattgctg cgacactgga gaacgacctc gctaggcttg
     2101 agaatgagaa cgcccgcctt gagaaggaca tagctaattt ggaacgcgat ttggccaaac
     2161 tggaacgcga agaggcatat tttctgcagg tcgacgaaca aaaactcatc tcagaagagg
     2221 atctgaatgc tgtgggccag gacacgcagg aggtcatcgt ggtgccacac tccttgccct
     2281 ttaaggtggt ggtgatctca gccatcctgg ccctggtggt gctcaccatc atctccctta
     2341 tcatcctcat catgctttgg cagaagaagc cacgttaacc caactttctt gtacaaagtg
     2401 gttgcaggaa agccaatccc aaacccactc ctaggtctcg acagtacagc atagaccggt
     2461 ccaccaccac caccaccact aaggatccgg ggttggggtt gcgccttttc caagcacctg
     2521 ctacatagcg gtacccctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2581 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2641 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2701 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2761 cttcgaattt aaatcggatc cagctgaatc taagtggatc tgcgatcgct ccggtgcccg
     2821 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2881 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2941 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     3001 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
     3061 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
     3121 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
     3181 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     3241 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
     3301 tctgcgccgt tacagatcta agctgtgacc ggcgcctact ctagagccgc catgtctaga
     3361 ctggacaaga gcaaagtcat aaactctgct ctggaattac tcaatggagt cggtatcgaa
     3421 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
     3481 cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat
     3541 catacccact cctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
     3601 aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
     3661 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
     3721 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
     3781 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
     3841 gattctatgc ccccacttct gaaacaagca attgagctgt tcgaccggca gggagccgaa
     3901 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
     3961 gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat
     4021 gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt
     4081 gacatgctcc ccggggaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     4141 cccggcccta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4201 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4261 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4321 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4381 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4441 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4501 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4561 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4621 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4681 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4741 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4801 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4861 tacaaggagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     4921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5521 aatcaacctc tggattacaa aatttgtggg taaacttgtt tattgcagct tataatggtt
     5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5641 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggaattga ctcaaatgat
     5701 gtcaattagt ctatcagaag ctcatctggt ctcccttccg ggggacaaga catccctgtt
     5761 taatatttaa acagcagtgt tcccaaactg ggttcttata tcccttgctc tggtcaacca
     5821 ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta aagaaacagt ggcagccagg
     5881 tttagccccg gaattgactg gattcctttt ttagggccca ttggtatggc tttttccccg
     5941 tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc
     6001 ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg
     6061 ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg
     6121 acgtaattac atccctgggg gctttggggg ggggctgtcc ctgatatcta taacaagaaa
     6181 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
     6241 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
     6301 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
     6361 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
     6421 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag
     6481 ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc
     6541 tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg
     6601 aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc
     6661 atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc
     6721 acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag
     6781 cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt
     6841 aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg
     6901 ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat
     6961 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7021 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7081 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7141 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7201 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7261 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
//
